How do bacteria and multicellular host organisms recognize and respond to each other? This is a fundamental question in studies of health and disease. Our project investigates the control of gene expression in a symbiosis between the alpha-proteobacterial species Sinorhizobium meliloti and its host, genus Medicago (such as alfalfa/M. sativa and barrel-medic/M. truncatula). Our lab has previously pioneered discoveries about host-bacterial signaling during the early steps of this complex symbiotic and developmental process. We have discovered that bacterial gene expression in symbiosis can follow one of six major patterns during symbiosis (that is, when and where the genes are turned on and off). In the next project period, we will identify promoters for these stage-specific genes and will determine the regulatory genes and proteins that control expression during development. One of the major players in gene expression control is the enzyme RNA polymerase, which recognizes DNA and reads out (expresses) only certain specific genes. In bacteria, a key component that determines which genes to express is the sigma (?) factor of this RNA polymerase. We found that ECF (extra-cytoplasmic function) ? factors are required for symbiosis: we now plan to analyze the mechanisms by which ECF ? factors are controlled in the context of a multi-stage developmental process. The bacterial stringent response usually occurs in reaction to nutrient deprivation; in Sinorhizobium, it is required for correct gene expression in symbiosis. We hypothesize this is programmed, and not merely a response to nutrient shift. We will directly test several hypotheses about how the stringent response is involved in the mechanism of differentiation. In parallel we will also look at how Sinorhizobium cells respond to alterations at the cell surface: This is important to understand because bacterial surface interactions with its eukaryotic host inevitably become an arena in which the two organisms signal and respond to each other.